JP2002363103A - Therapeutic use of selective pde10 inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for treating a certain kind of neurologic and psychiatric disorders in mammals including humans, especially a method for treating mood, movement and anxiety disorders, schizophrenia, drug, e.g. alcohol addiction and disorders having deficient cognitive power as expression of symptoms. SOLUTION: This method for treating comprises administering a selective phosphodiesterase(PDE)10 inhibitor. Furthermore, the method includes a method for treating by use of papaverine as a selective inhibitor of PDE10.

Description

DETAILED DESCRIPTION OF THE INVENTION

BACKGROUND OF THE INVENTION The present invention relates to the treatment of disorders of the central nervous system. More particularly, the present invention relates to the treatment of neurological and psychiatric disorders, such as psychiatric disorders and disorders including cognitive deficits as symptoms. The invention further relates to inhibiting PDE10.

[0002] Cyclic nucleotides, cyclic-adenosine monophosphate (cAMP) and cyclic-guanosine monophosphate (cGMP), are intracellular proteins that regulate a vast array of intracellular processes, including neurons of the central nervous system. Acts as a second messenger. cAMP is synthesized by a family of membrane-bound enzymes, adenylyl cyclase. A broad range of serpin family receptors regulates these enzymes by a coupling mechanism mediated by heterotrimeric G-proteins. Increased intracellular cAMP leads to activation of cAMP-dependent protein kinase, which regulates the activation of other signaling kinases, transcription factors, and enzymes by their phosphorylation. Cyclic AMP is
It also directly affects the activity of ion channels, phosphodiesterases, or guanine nucleotide exchange factors that are regulated by cyclic nucleotides. Recent studies further suggest that intracellular cAMP can function as a precursor to the neuromodulator, adenosine, following transport from that cell.

[0003] Guanylyl cyclase, which synthesizes cGMP, exists in membrane-bound and cytoplasmic forms. The membrane-bound type is A
Like NP (atrial natriuretic peptide), it couples to receptors linked to G-proteins, while soluble guanylyl cyclase is activated by nitric oxide (Wang, X. and Robinso).
n, P. J. Journal of Neuroche
mystery 68 (2): 443-456,199
7). Like cAMP, downstream mediators of cGMP signaling in the central nervous system are cGMP-gated ion channels, cGMP-regulated phosphodiesterase and cGMP.
Includes dependent protein kinases. The cyclic nucleotide is
Because they have an important role in signal transduction within the central nervous system, the use of compounds that affect the modulation of cyclic nucleotide signaling provides therapeutic benefit.

A major mechanism for regulating cyclic nucleotide signaling is through phosphodiesterase-catalyzed catabolism of cyclic nucleotides. There are 11 known families of phosphodiesterases (PDEs) encoded by 21 different genes. Each gene typically provides multiple splice variants that further contribute to isozyme diversity. The family of PDEs includes cyclic nucleotide substrate specificity, regulatory mechanisms,
And functionally based on the sensitivity of the inhibition. Furthermore,
PDEs are specifically expressed in organs, including in the central nervous system. As a result of these unique enzyme activities and localization,
Different PDE isozymes can perform unique physiological functions. In addition, compounds that can selectively inhibit unique PDE families or isozymes can provide special therapeutic effects, reduced side effects, or both.

[0005] PDE10 is identified as a unique family based on its first amino acid sequence and unique enzymatic activity. Homology screening of the EST database identified the mouse PDE10A with the phosphodiesterase PDE10.
Revealed as the first member of the family (Fuji
Shige et al. , J. et al. Biol. Chem.
274: 18438-18445, 1999;
hney, K .; et al. , Gene 234: 10
9-117, 1999). Mouse homologues have been further cloned (Soderling, S. et al., P.
rc. Natl. Acad. Sci. USA 96:
7071-7076, 1999), and N-terminal splice variants of both rat and human genes have been identified (Kotera, J. et al., Bioche).
m. Biophys. Res. Comm. 261: 55
1-557, 1999; Fujishige, K .; et
al. , Eur. J. Biochem. 266: 11
18-1127, 1999). There is a high degree of homology between species. Mouse PDE10A1 is a 779 amino acid protein that contains cAMP and cAMP.
GMP is hydrolyzed to AMP and GMP, respectively. P
Affinity of DE10 for cAMP (K _m = 0.05 μm
M) is higher than that for cGMP (K _m = 3 μM). However, schematic 5 fold greater V _max of cGMP than for cAMP leads to the suggestion that PDE10 is a unique cAMP inhibition cGMP-ase (Fujish
image et al. , J. et al. Biol. Chem. 27
4: 18438-18445, 1999).

[0006] PDE10 is also uniquely located in mammals compared to other PDE families. MRNA for PDE is highly expressed only in testis and brain (Fu
Jishige, K .; et al. , Eur J Bi
ochem. 266: 1118-1127, 1999;
Soderling, S.M. et al. Proc. N
atl. Acad. Sci. 96: 7071-707
6, 1999; Loughney, K .; et al. ,
Gene 234: 109-117, 1999). These first studies showed that PDE10 expression in the brain was highest in the striatum (caudate and putamen), nucleus accumbens, and olfactory tubercle. More recently, a detailed analysis of the expression pattern of PDE10 mRNA in rodent brains has been performed (Seeger, TF et a.
l. , Abst. Soc. Neurosci. 26: 3
45.10, 2000).

SUMMARY OF THE INVENTION The present invention provides a method of treating anxiety or psychiatric disorders in a mammal, including a human, comprising administering to said mammal an amount effective in treating said anxiety or psychiatric disorder. Administering a selective PDE10 inhibitor.

[0008] The present invention further provides a method of treating anxiety or psychiatric disorders in a mammal, including a human, comprising providing the mammal with an effective amount of a selective PDE10 inhibitor in inhibiting PDE10. Administration.

Examples of psychiatric disorders that can be treated according to the present invention include, but are not limited to, paranoid, schizophrenic, tense, undifferentiated, or residual schizophrenia; schizophrenic disorders Substance-induced mental disorders, such as alcohol, amphetamine, timer, cocaine, hallucinogens, inhalants, opioids, or phencyclidine; paranoia; paranoid or depressed schizophrenic disorders; paranoid disorders; Pathological personality disorders; and schizophrenic personality disorders.

[0010] Examples of anxiety disorders that can be treated according to the present invention include, but are not limited to, panic disorder; agoraphobia; specific phobia; social phobia; obsessive-compulsive disorder; Disorders; acute stress disorders; and general anxiety disorders.

[0011] The present invention further provides a method for treating a movement disorder selected from Huntington's disease and movement disorder associated with dopamine agonist treatment in mammals including humans,
The method comprises administering to the mammal an effective amount of a selective PDE10 inhibitor in treating the disorder.

The present invention further provides a method of treating a movement disorder selected from Huntington's disease and movement disorder associated with dopamine agonist treatment in mammals, including humans,
The method comprises administering to the mammal an effective amount of a selective PDE10 inhibitor in inhibiting PDE10.

[0013] The present invention further provides a method of treating a movement disorder selected from Parkinson's disease and lower limb anxiety syndrome in a mammal, including a human, and selecting an effective amount in treating the disorder in the mammal. Administering a selective PDE10 inhibitor.

[0014] The present invention further provides a method for treating a movement disorder selected from Parkinson's disease and lower limb anxiety syndrome in a mammal, including a human, wherein the mammal is provided with a selective amount of a PDE10 effective to inhibit PDE10. Administering a PDE10 inhibitor.

The present invention further relates to drug addiction in mammals, including humans, such as alcohol, amphetamine, cocaine,
Alternatively, there is provided a method of treating opiate drug addiction, comprising administering to said mammal an amount of a selective PDE10 inhibitor effective in treating drug addiction.

The present invention further relates to drug addiction in mammals, including humans, such as alcohol, amphetamine, cocaine,
Alternatively, there is provided a method of treating opiate addiction, comprising administering to said mammal an amount of a selective PDE10 inhibitor effective in inhibiting PDE10.

As used herein, "drug addiction"
It refers to an abnormal desire for a drug, and is generally characterized by impaired motivational phenomena, such as the manifestation of compulsive and intense drug cravings for taking the desired drug.

The present invention further provides a method of treating a disorder, including cognitive deficits in a mammal, including a human, as a symptom, the method being effective in treating the mammal with cognitive deficits. Administering an amount of a selective PDE10 inhibitor.

The present invention further provides a method of treating a disorder, including cognitive deficits as a symptom, in a mammal, including a human, comprising administering to said mammal an effective amount of a PDE10 inhibitor. Administering a selective PDE10 inhibitor.

[0020] The phrase "lack of cognition" as used herein in "disorders including loss of cognition as a symptom" refers to a particular individual's other individuals within the same general age group. A subnormal function in one or more aspects of cognition, such as memory, intelligence, or learning and logical ability, as compared to. “Lack of cognition” further refers to a decline in the function of any particular individual in one or more aspects of cognition, for example, as occurs as age-related cognitive decline.

Examples of disorders that can be treated according to the present invention, including cognitive deficits as symptoms are dementia, such as Alzheimer's disease, multiple infarct dementia, alcoholic dementia or other drug-related dementia, intracranial tumors Or dementia associated with brain trauma, dementia associated with Huntington's disease or Parkinson's disease, or AIDS-related dementia; delirium, amnestic disorder; post-traumatic stress disorder; mental retardation; Power impairment, or impaired written expression; attention deficit / hyperfunction; and age-related cognitive decline.

The present invention further provides a method of treating a mood disorder or a mood disorder in a mammal, including a human, and selecting an effective amount in treating the mammal in the mammal. Administering a selective PDE10 inhibitor.

The present invention further provides a method of treating a mood disorder or a mood disorder in a mammal, including a human, wherein the mammal is provided with an effective amount of a selective PDE10 inhibitor in inhibiting PDE10. Administration.

Examples of mood disorders or mood symptoms that can be treated according to the present invention are not limited, but the mood disorders or mood symptoms may be of mild, moderate or severe type. Onset of depressive symptoms, onset of manic or mixed mood symptoms, onset of hypomanic mood symptoms; onset of atypical signs of depressive mood symptoms; onset of depressive signs of depressive symptoms; Onset of depressive symptoms of pathological signs; onset of mood symptoms starting after parturition; depression after seizures; severe depressive disorder; dysthymia disorder; mild depressive disorder; premenstrual discomfort disorder; Schizophrenic post-psychotic depressive disorder; severe depressive disorder overlapping with psychotic disorder such as delusional disorder or schizophrenia; manic depressive disorder, eg, manic depressive I disorder, manic depressive II disorder, and circulation Including sexual disorders.

As used herein, "selective PDE1"
The term "zero inhibitor" refers to a substance, such as an organic molecule, which refers to any other PDE enzyme, in particular any PDE1
It effectively inhibits enzymes from the PDE10 family to a greater extent than enzymes from the -9 family or any PDE11 enzyme. In one embodiment, the selective PDE
10 Inhibitors are substances, such as organic molecules, which have a K value that has the effect of inhibiting any other PDE enzyme.
_i have a K _i for PDE10 inhibition of about one-tenth or less. In other words, the substance inhibits PDE10 activity to the same extent at concentrations about one-tenth or less than those required for any other enzyme.

Generally, the substance will have an IC of about 10 μM or less, preferably about 0.1 μM or less.
Having a value of ₅₀ would effectively inhibit PDE10 activity.

"Selective PDE10 inhibitors" include, for example, P
The ability of a substance to inhibit DE10 activity can be identified by comparing it to its ability to inhibit PDE enzymes from other PDE families. For example, the substance
PDE10 activity and PDE1, PDE2, PDE3
A, PDE4A, PDE4B, PDE4C, PDE4
D, PDE5, PDE6, PDE7, PDE8, PDE
9, and its ability to inhibit PDE11.

In one embodiment of the treatment method of the invention described above, the selective PDE10 inhibitor is papaverine. The present invention further relates to mammalian PDE10s, including humans.
Providing a method of selectively inhibiting
Administering an effective amount of papaverine in inhibiting E10.

The term "treatment" in the term "method of treating a disorder" refers to reversing, alleviating or inhibiting the progression of the disorder to which such term applies, or the manifestation of one or more disorders. Point to. As used herein, the term further includes prevention of the onset of the disorder or the manifestation of any symptoms associated therewith, depending on the condition of the patient, preventing the disorder, and preventing or preventing the disorder or initiation. It also includes reducing the severity of any of the previous episodes of the symptom. "Treatment" as used herein also refers to preventing the recurrence of a disorder.

For example, “treatment of schizophrenia or schizophrenia or schizophrenic affective disorder” as used herein further refers to the manifestation of one or more symptoms (positive, negative, and other) of said disorder. For example, treatment of concomitant delusions and / or hallucinations. Schizophrenia,
And the manifestations of schizophrenia and schizophrenia affective disorder can be disordered speech, flattening of emotions, oligodrome, discomfort, inappropriate emotions, unpleasant moods (eg, in the form of depression, anxiety or anger) ), And some signs of cognitive dysfunction.

“Mammal” as used herein refers to
It refers to any member of the class "Mammalia" and includes, but is not limited to, humans, dogs, and cats. DETAILED DESCRIPTION OF THE INVENTION Isozymes of PDE2, 3, and 5, including human PDEs, can be prepared, for example, from the corpus cavernosum; isozymes of PDE1, including humans, from the ventricle; and isozymes of PDE4, including humans, can be prepared from skeletal muscle. it can. PDE6
Can be prepared, for example, from the canine retina. A description of the preparation of enzymes from natural tissues can be found, for example, in Boolell, M .; et
al. , Int. J. Impotence Rese
arch 8: 7-52, 1996.

PDE7-11 can also be prepared from natural tissues. Isozymes from the PDE7-9 and 11 families can alternatively be used as Fish
er, D .; A. et al. , Biochem. Bio
phys. Res. Comm. 246,570-57
7, 1998; Soderling, S .; H. et a
l. , PNAS 96: 7071-7076,199.
9; Fisher, D .; A. et al. , J. et al. Bio
l. Chem. 273, 15559-15564, 19
98b; and Fawcett, L .; , Et al. , P
For example, as described in NAS 97: 3702-3707, 2000, it can be generated from a full-length human recombinant clone transfected into SF9 cells. PDE10 can also be generated from recombinant rat clones transfected into SF9 cells (Fujishige, et al.,
European Journal of Bioch
emistry, Vol. 266,1118-1127
(1999)). The enzyme is then prepared by FPLC from the soluble fraction of the cell lysate as described for PDE6. The references mentioned above are hereby incorporated by reference in their entirety.

In one assay, the substance is PDE
Screened for inhibition of cyclic nucleotide hydrolysis by PDEs from 10 and other gene families. The concentration of cyclic nucleotide substrate used in each individual PDE assay was 1 _Km concentration.
/ 3, allowing comparison of IC ₅₀ values between different enzymes. PDE activity was determined as previously described (Fawcette).
t al. , 2000) using a method based on scintillation proximity analysis (SPA).
The effect of the PDE inhibitor is that a certain amount of enzyme (PDE1-1
1) is determined by assaying in the presence of varying concentrations of the substance and low concentrations of substrate such that the IC ₅₀ approaches K _i (at a concentration of 1/3 K _m , unlabeled and [ ³ H] cGMP or cAMP with a 3: 1 label ratio).
The volume of the final assay Assay buffer [20 mM of Tris-HCl, MgCl ₂ of pH 7.4, 5 mM,
1 mg / ml bovine serum albumin]. The reaction is initiated with the enzyme and incubated at 30 ° C. for 30-60 minutes to obtain <30% substrate conversion,
Then, 50 μl of yttrium silicate SPA beads (A
mersham) (containing 3 mM unlabeled cyclic nucleotides for PDEs 9 and 11, respectively). The plate is resealed and shaken for 20 minutes, after which the beads are allowed to settle in the dark for 30 minutes and then topcount plate reader (Packa
rd, Meriden, CT). Radioactive units can be converted to percent activity of the uninhibited control (100%), plotted against inhibitor concentration, and the IC ₅₀ value of the inhibitor can be calculated using the Microsoft Exc
El can be obtained using the 'Fit Curve' extension.

One example of a selective PDE10 inhibitor is papaverine (1-[(3,4-dimethoxyphenyl) methyl] -6,7-dimethoxyisoquinoline). Papaverine is a known effective smooth muscle relaxant used in the treatment of cerebral and coronary vasospasm, and diastolic dysfunction. Although the basis of their therapeutic activity is not fully understood, they generally result from the activity of papaverine as a non-selective phosphodiesterase inhibitor (The The
Pharmacological Basisof Th
erapeutics; Sixth Edition;
A. G. FIG. Gilman, L .; S. Goodman, A .;
Gilman (eds.) Macmillan Pub
living Co. , New York, 198.
0, p. 830). Papaverine is a naturally occurring plant alkaloid, but its complete biosynthesis, for example,
Brochm, incorporated herein by reference.
anna-Hanssen et al. , J. et al. Phar
m. Sci. 60: 1672,1971.

The selective PDE10 inhibitors can be administered in accordance with the present invention, either alone or in combination with pharmaceutically acceptable carriers, in either single or multiple doses. Suitable pharmaceutical carriers include inert solid diluents or fillers, sterile aqueous solution and various organic solvents. The pharmaceutical composition thus formed can then be easily administered in various dosage forms such as tablets, powders, lozenges, syrups, solutions for injection and the like. These pharmaceutical compositions can, if desired, contain additional ingredients such as fragrances, binders, excipients, and the like. Thus, for oral administration purposes, tablets containing various excipients, such as sodium citrate, calcium carbonate, and calcium phosphate, may have various disintegration properties, such as starch, methylcellulose, alginic acid and certain complex silicates. With the agent, it can be used with binders such as polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often useful for tableting purposes. Solid compositions of a similar type can also be used as fillers in soft and hard filled gelatin capsules. Preferred materials for this include lactose or milk sugar and high molecular weight polyethylene glycols. If an aqueous suspension or elixir is desired for oral administration, the essential active ingredients therein are various sweetening or flavoring agents, coloring substances or dyes and, if desired, emulsifying or suspending agents. When,
It can be mixed with diluents such as water, ethanol, propylene glycol, glycerin and combinations thereof.

For parenteral administration, solutions containing the selective PDE10 inhibitor in sesame or peanut oil, aqueous propylene glycol, or sterile aqueous solution may be employed. Such aqueous solutions should be suitably buffered if necessary, and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. The sterile aqueous media used are all readily available by standard techniques known to those skilled in the art.

The selective PDE10 inhibitor may be administered orally, transdermally (eg, by using a patch), parenterally (eg, intravenously), rectally, or topically in the treatment method of the present invention. Can be. Generally, a PDE for treating a disorder or condition by the methods described herein.
The daily dose of 10 inhibitors is generally about 0.01
To about 100 mg / kg of the patient's body weight to be treated. As an example, a selective PDE10 inhibitor may have an average body weight (about 70
kg) of a human adult from about 1 mg to about 7 mg per day.
It may be administered in a single or divided (i.e., multiple) dose of up to 000 mg, preferably in the range of about 1 mg to about 1000 mg per day. Changes based on the dosage ranges described above may be based on the weight of the patient being treated,
It can be done by a physician of ordinary skill in view of known conditions such as age and condition, degree of distress, and the particular route of administration chosen.

The following examples illustrate the invention. However, it is to be understood that the invention as fully described herein and as claimed, is not intended to be limited by the details of the following examples.

Embodiment Embodiment 1 Selective PDE10 inhibitors: Papaverine: Papaverine was screened for inhibition of hydrolysis of cyclic nucleotides by PDE10 and a group of PDEs from other gene families. Each individual PDE
The concentration of cyclic nucleotide substrate used in the assay was
It was 1/3 of the Km concentration. This allowed comparison IC ₅₀ values between the different enzymes.

PDE activity was measured using the yttrium silicate SPA beads assay described above in the Detailed Description section. Radioactive units were compared to the uninhibited control (10
0%), plotted against the concentration of inhibitor, and the IC ₅₀ value of the inhibitor
Obtained using the soft Excel 'Fit Curve' extension.

The papaverine had an IC _{50 of} 18 nM (Table 1).
It was observed to be an exceptionally potent, competitive inhibitor of PDE10 with value. Papaverine was not very potent against all other PDEs tested.
Following PDElO, most potently inhibited enzyme by papaverine was PDE4D with 19-fold lower 320 nM IC ₅₀ than that for PDElO. Therefore, these data indicate that papaverine is a selective PDE10
It is for the first time demonstrated that this compound is an inhibitor and can be used in the study of the physiology of this enzyme.

Table 1. The IC ₅₀ values for papaverine inhibition of PDE described. The IC ₅₀ is, for each enzyme,
It was measured at a substrate concentration of 1/3 of the Km value, allowing a comparison between the enzymes. The PDE10 selectivity ratio is given for a given PD.
The IC ₅₀ values for E is obtained by dividing an IC ₅₀ value for PDElO.

[0043]

[Table 1] Embodiment 2. FIG. Medium spiny neuron (Mediu
m Spiny Neuron)
Effect of selective PDE10 inhibitor on ptide metabolism: The effect of the selective PDE10 inhibitor papaverine, as measured in Example 1, was tested in primary cultures on metabolism of cyclic nucleotides in rat medium spiny neurons did.

Neurons cultured in the presence of BDNF from the embryonic striatum of E17 rats exhibited a phenotype very similar to that previously described (Ventimiglia et al.).
al. , Eur. J. Neurosci. 7 (199
5) 213-222). Approximately 50% of these neurons stained positive for GABA immunoreactivity, confirming the presence of medium spiny neurons in culture.
The message of PDE10 expression in these cultures at 5 DIV was confirmed by an RNase protection assay.

Striatum cultures were prepared as previously described (Ventimiglia et al., Eu).
r. J. Neurosci. 7: 213-222, 19
95). Briefly, excised striatum (caudal nucleus and putamen) from E17 rats were isolated to produce single cell suspensions and poly- at a density of 5 × 10 ⁴ neurons / well. Placed in a multi-well plate coated with L-ornithine / laminin. Cells were supplemented with B27 supplement and BDNF (100
ng / mL) in Neurobasal medium. Experiments were typically performed after 4 days in vitro. Medium spiny neurons make up the major cells of these cultures (50-60%, GA
BA immunoreactivity).

In the RNase protection assay, the RNA
From primary cultures of rat medium spiny neurons prior to (Iredale, PA, et al., Mo.
l. Pharmacol. 50: 1103-1110,
1996) with a gradient of 5.7M cesium chloride at 150,000 xg by centrifugation at 20 ° C for 21 hours. Transfer the RNA pellet to pH
5.2 Resuspended in 0.3 M sodium acetate, precipitated in ethanol and the concentration was determined by spectrophotometry. PDE10 riboprobe
e) was prepared by PCR amplification of a 914 bp fragment (corresponding to bp 380-bp 1294) isolated from mouse cDNA. This fragment was then used to construct pGEN3Zf
Cloned. The vector was linearized and T7
[ ³² P] -labeled antisense riboprobes were synthesized using RNA polymerase. RNase protection assays were performed using the RPAII kit (Ambion). Briefly, 5 μg of total cellular RNA was combined with [ ³² P] -labeled PDE10 riboprobe (about 105 cpm / sample),
Hybridized overnight at 42 ° C. The next day, the sample was
Incubate withases A and T1 for 30 minutes at 37 ° C., and then precipitate the protected double stranded RNA fragment and run on a 6% polyacrylamide gel containing urea.

To analyze the effect of papaverine on cyclic nucleotides, striatal cell cultures after 4 days in vitro were washed with Ca ²⁺ / Mg ⁺ free phosphate buffered saline and Phosphate buffered saline without Ca ²⁺ / Mg ⁺ , 30 mM HEPES, 1 m
Preincubation was performed for 1 hour in a buffer containing M CaCl ₂ , 1 mg / mL dextrose, and 5 mM MgCl ₂ . Striatal cells were exposed to a phosphodiesterase inhibitor and incubated at 37 degrees Celsius for 20 minutes. When measuring cGMP, neurons were stimulated for 2 minutes with a nitric oxide source, sodium nitroprusside, following a 20 minute incubation with the compound. When measuring cAMP, neurons were stimulated with forskolin, an activator of adenylate cyclase, during a 20 minute incubation with compound. Cells were purified using a 9: 1 mixture of cAMP SPA direct screening assay buffer (0.05 M acetate and 0.01% sodium azide) and buffer A (133 mg / mL dodecyltrimethylammonium bromide). To dissolve and freeze the lysate on dry ice. cG
MP [I125] or cAMP [I125] scintillation proximity assay (SPA) systems (Amersha, respectively)
m code RPA 540 and RPA 559) were used to detect the concentration of each cyclic nucleotide in the cell lysate.

With papaverine alone, c in striatal cultures
It did not affect low basal concentrations of either AMP or cGMP. Therefore, the effect of cAMP or cGMP synthesis under conditions stimulated by forskolin or the NO donor sodium nitroprusside (SNP), respectively, was studied. Forskolin (0.1-10μ) for 20 minutes
Stimulation of the culture with M) showed a concentration-dependent increase in cAMP concentration. Similarly, the SNP of the culture (3-1000
(M) for 2 minutes showed a concentration-dependent increase in cGMP concentration. Forskolin alone (10
μM) does not change the concentration of cGMP and
(300 μM) also did not increase cAMP concentration.
To determine the effect of papaverine on cAMP and cGMP metabolism, striatal cultures were incubated with various concentrations of compound and then sub-maximally effective concentrations of either forskolin (1 μM) or SNP (100 μM). Stimulated. These concentrations of forskolin or SNP caused a 2-3 fold increase over the bases of cAMP and cGMP, respectively. Papaverine had an EC ₂₀₀ (21.7 μM) of 11.7 μM in SNP-induced cGMP accumulation.
A concentration-dependent increase with the concentration of the inhibitor giving a two-fold increase) values occurred (Table 2). The greatest effect was observed at 100 μM, at which point the cGMP concentration was increased 5-fold over that of the culture stimulated with SNP alone. Papaverine also caused an increase in cAMP accumulation in forskolin-stimulated cultures. However, the compound is cAMP
Was 3.3 times less potent than cGMP in promoting the increase in The effect of papaverine in striatal cultures was compared to other PDE inhibitors with different selectivities (Table 2). IBMX, a non-selective inhibitor, is SNP-
Alternatively, a concentration-dependent (3-100 μM) increase occurred in both cGMP and cAMP accumulation with EC _{200 of} 19 and 30 μM, respectively, in forskolin-stimulated cultures. The selective PDE4 inhibitor rolipram
m) increased the accumulation of forskolin-stimulated cAMP with an EC ₂₀₀ value of 2.5 μM and required a 10-fold higher concentration to double the rate of cGMP accumulation. PDE
Zaprinast (za, a cGMP inhibitor)
prinast) doubled the cAMP concentration in these neurons at a concentration of 98 μM. However, 100 μM of this compound did not completely double the concentration of cGMP. These data indicate that papaverine has a unique effect on the regulation of cyclic nucleotides in medium spiny neurons, and that
It is shown for the first time that it is due to the selectivity for E10.

Table 2. EC ₂₀₀ values for elevation of cGMP or cAMP during primary culture of rat striatal neurons.
EC ₂₀₀ values refer to the concentration that produces a 200% increase in cGMP or cAMP in SNP- or forskolin stimulated cultures, respectively. Each value is the mean ± SEM from the indicated number of experiments (n). E. FIG. M. It is. Each experiment was repeated with 3-6 sister cultures.

[0050]

[Table 2] Embodiment 3 FIG. Selectivity in animal models of basilar ganglion function
Effects of PDE10 inhibitors: Studies in human and non-human mammals show that basilar ganglia modulate a range of motility and cognitive and emotional / appetite behaviors (G
raybiel, A .; M. Current Biolo
gy 10 (14): R509-11, 2000). A murine experimental model has been developed that can be used to evaluate the effects of compounds on basilar ganglion function. Papaverine is found to have unexpected and unique aspects of behavioral effects in these two models.

The effect of papaverine alone and in combination with haloperidol was tested on its ability to induce catalepsy in male CD rats. This animal model was used to analyze the effects of compounds on basilar ganglion output. Papaverine (1.0, 3.2, 10,
Or 32 mg / kg) or vehicle was administered subcutaneously. In some experiments, this was followed immediately by 1.
Haloperidol at 0 mg / kg (in 0.3% sc tartaric acid) was administered. Thirty minutes after drug administration, the degree of catalepsy was increased by lifting the rat's front paws (10c
m) Latency on the rod (1 cm diameter) and removal of both forepaws from the rod was quantified by measuring the latency truncation as 30 seconds. Latency is Krusk
For comparison by all-Wallace analysis of variance,
Each treatment group was classified. Post-hoc analysis was performed by Mann
According to the Whitney U test.

The antipsychotic haloperidol was described previously (Chartoff, E. et al., J Phar).
macol. Exp. Ther. 291: 531-53
7, 1999), this model produces strong catalepsy. The maximum effective dose of haloperidol is 1 mg / kg, s. c. Was found.
In contrast, papaverine alone at 32 mg / kg, s.
c. When administered at doses up to (p = 0.86),
It did not induce catalepsy. However, as shown in FIG. 1, papaverine increased the catalepsy effect of submaximal (0.32 mg / kg, sc) doses of haloperidol (p <0.001). The minimum effective dose of papaverine for haloperidol-induced increase in catalepsy is 3.2 mg / kg, s. c. It is. This experiment demonstrated that papaverine could alter basilar ganglion output in a direction consistent with antipsychotic activity.

Embodiment 4 FIG . Animal models for mental illness
Effect of Selective PDE10 Inhibitor on Papaverine Next, the effect of papaverine on rat locomotor activity measured in a shuttle box was studied. The decrease in PCP-stimulated locomotor activity in mice has been accepted as a primary screen in search of new antipsychotics. Newer atypical antipsychotics generally have a P-value for amphetamine-stimulated motor activity.
Demonstrates its selective inhibition of CP. Mature male S
prague-Dawley rats (250-300
g) is replaced by Charles River (Wilming).
ton, MA). Locomotor activity was measured using a commercially available shuttle box (Coulbourn In
instruments, Allentown, PA). After drug administration, data was collected at 5 minute intervals for 1 hour. Rats receive vehicle (5%
DMSO, 5% Emulphor,
90% physiological saline), phencyclidine (PCP, Si
gma Chem. Co. )) Or amphetamine sulfate (RBI), followed immediately by either vehicle or test compound. Statistical analysis is t-
Performed using a test.

Both the psychostimulants amphetamine and phencyclidine (PCP) resulted in a strong increase in motor activity in this model. Papaverine alone (32mg / k
g, i. p. ) Resulted in a slight decrease in locomotor activity, which was statistically significant in one study (FIG. 2). However, this same dose of papaverine provides a behaviorally equivalent dose of amphetamine (1 mg / kg, ip).
Without affecting that caused by 3.2 mg /
kg, i. p. Caused a significant decrease in locomotor activity stimulated by phencyclidine.

[0055] As described in the study destination, using the in situ hybridization, PDE in the striatum, nucleus accumbens and olfactory tubercle
High expression of 10 mRNAs has already been reported (S
eeger, T .; F. Et al. , Supra). Applicants have further found using monoclonal antibodies against the PDE10 protein, a correspondingly high concentration of the PDE10 protein in these brain regions. Applicants have found PDE10 mRNA to be expressed at high levels in medium spiny neurons in the striatum and nucleus accumbens. Medium spiny neurons are the output neurons of the striatum, nucleus accumbens, and olfactory tuberculosis; and are about 95% of all neurons in these brain structures. In addition, high concentrations of PDE10 protein are observed in the processes (axons and terminals) of medium spiny neurons that protrude from the striatum, nucleus accumbens, and other cerebral areas including the pallidum and substantia nigra from the olfactory tubercle. Was. These latter brain regions have low or undetectable concentrations of PDE1 themselves.
It has 0 mRNA. Therefore, the PDEs in these areas
The high concentration of 10 is due to the axons and terminals of medium spiny neurons. In addition, Applicants' work has demonstrated PDE10 mRNA and protein expressed at low concentrations in other brain regions, namely cortical, hippocampal and cerebellar neurons.

High concentrations of PD in the striatum and nucleus accumbens
The expression of E10 is of particular interest given that these are the major cortical input nuclei of the basilar ganglia, as well as the major terminal regions for dopaminergic processes in the midbrain. The striatum and its ventral extension, the nucleus accumbens, receive glutamate-active afferents from substantially each region of the cerebral cortex and serve as hypocortical anabolic sites of extensive cortical activity. I do. The dorsal striatum is generally involved in the regulation of motor behavior, while the ventral region, including the locus, is involved in functions in regulating emotional / appetite behavior. Thus, Applicants believe that PDE10 may be involved in signaling pathways that regulate many of these fundamental physiological processes.

The cortical input to the striatum is
GABAergic midi, which makes up 95% of neurons
Primary to Amespiny process neurons (MSN)
Provides irritating propulsion. Glutamic acid action of MSN
Activation, which in turn causes large amounts of dopaminergic activity from the midbrain
Adjusted by input. These two afferent systems
The nature of antagonist nature has been proven in many studies.
ing. For example, exercise stimulant activity in laboratory animals
Is the dopa of the NMDA subtype of glutamate receptor
Min receptor agonist or antagonist
(Carlsson, ML.
and Carlsson, A .; Trend New
osci. 13: 272-276, 1990). Halope
D like a ridor_TwoDopamine receptor antagonist
Cataleptic effect of the haloperidol-induced gene
As with expression, by NMDA receptor antagonists
Reduced (Chartoff, E et al., J.
 Pharmacol. Exp. Ther. 291: 5
31-537, 1999). More recently, D _TwoDo
-Blocking of papamine receptors is associated with striatal NMDA receptor
Will increase sphorylation or activation state
(Leveque et al., Jo)
urnal of Neuroscience 20
(11): 4011-4020, 2000).

The recognition that all clinically effective antipsychotics possess potent D ₂ antagonist activity suggests that the manifestation of schizophrenia is a consequence of the hyperactivity of the dopamine system in the central limbic system. Leads to the original hypothesis that The evaluation of the ability of chemical compounds to reduce the stimulating properties of direct or indirect dopamine agonists has become an important laboratory test in the quest for new antipsychotics. More recently, the ability of NMDA receptor antagonists such as PCP to faithfully mimic the onset of positive, negative and cognitive symptoms of human schizophrenia (Luby et al., 1).
959; Rosenbaum et al, 1959;
Krystal et al. 1994) led to the development of a hypothetical theory of schizophrenia. Briefly, this hypothesis states that inhibition of striatal-mediated behavior is inadequate in schizophrenia, as a result of glutamatergic and specifically NMDA receptor-mediated neurotransmitter depletion. It is proposed that. This hypothesis, assuming the their ability to directly or indirectly disinhibition cortical input (as described above) for the striatum, complete with known anti-neuropathic effects of D ₂ dopamine receptor antagonists Matches. The fidelity with which PCP replicates the manifestations of human schizophrenia has led to the use of murine PCP-stimulated locomotion as a primary screen in the search for novel antipsychotics. Evidence that newer, and perhaps more potent, atypical antipsychotics show selective activity against that of PCP over that of amphetamine-stimulated motor activity
It is clear that this method is supported (Gl
eason S.A. D. and Shannon H. et al. E. FIG.
Psychopharmacol. 129: 79-8
4, 1997).

The current methods of antipsychotic treatment are:
While generally targeting membrane receptors, Applicants have
Increase NMDA receptors in SN or dopamine D ₂
It is proposed herein that intracellular manipulation of PDE10, which reduces receptor effects, can further produce antipsychotic effects. Increased cAMP and cGMP activities are known to enhance the response of striatal neurons to glutamate agonists, including NMDA (Colwell, CS and MS Levi).
ne, J Neuroscience 15 (3) 17
04-1713, 1995). Haloperidol's neuroleptic effect was further enhanced by cAMP levels (Ward, RP an.
d D. M. Dorsa, Neuroscience
89 (3): 927-938, 1999) and activation of PKA (Adams, MR et al., Proc.
Natl Acad Sci USA 94: 1215
7-12161, 1997). Striatum cG
MP levels were further increased after blockade of the D ₂ receptor (Alt
ar, C.E. A. et al. , Eur J .; Pharm
acol. 181: 17-21, 1990) and P
KG is an endogenous inhibitor of protein phosphatase I, D
It is known to phosphorylate some of the same downstream substrates, such as PKA, including ARP (Green)
guard Pet al. , Brain Res. Re
v. 26: 274-284, 1998). Accordingly, Applicants believe that an agent capable of selectively increasing the concentration of cyclic nucleotides in MSN in the striatum would increase striatum function with the resulting antipsychotic effect. Can be reasonably expected, and PDE10 inhibitors show that such compounds are capable of cAMP and cGMP PD
The metabolism of the E10 catalyst is determined by the MS of these cyclic nucleotides.
The hypothesis is that it has therapeutic efficacy in treating mental illness because it increases and inhibits its concentration in N.

Experiments with papaverine described above demonstrate that selective PDE10 inhibitors can increase the concentration of cyclic nucleotides in MSN in the striatum of basilar ganglia. Studies with the animal models described above indicate that selective PDE10 inhibitors can produce unique and unexpected changes in basilar ganglion function.

In addition to mental illness, the abnormal function of the basilar ganglia is dementia / hyperactivity disorder (ADHD) as well as related attention disorders (Seeman, P. et al., M.
molecular Psychiatry 3: 386
-96, 1998), depression (Kapur, S., Bio).
l. Psychiatry 32: 1-17,199
2; Willner, P .; , Brain Res. 28
7: 225-236, 1983), and substance abuse (Se
If, D. W. Annals of Med. 30: 3
79-389, 1998). Parkinson's disease, lower limb anxiety syndrome (H
ening, W.M. et al. , Sleep 22: 9
70-999, 1999) and several neurological disorders, including Huntington's disease, have further been linked to basilar ganglion dysfunction. Thus, based on Applicants' work described herein, Applicants believe that the PDE10 inhibitor
We believe that these disorders have therapeutic impact.

As disclosed above, the Applicants have
We found that E10 mRNA and protein are also expressed in hippocampal and cortical neurons. Since the process of cognition depends on hippocampal and cortical functions, Applicants
DE10 also plays a role in the process of cognition, and PDE10 inhibitors are used to treat disorders with characteristic components of poor cognitive function, such as Alzheimer's disease and age-related cognitive decline (ARCD). Believe that it can be used against.

[Brief description of the drawings]

FIG. 1 is a bar graph showing animal catalepsy versus increasing doses of papaverine. Gray bars indicate papaverine in combination with haloperidol and of haloperidol-induced catalepsy
The synergistic effect of papaverine is shown. Black bars indicate papaverine alone. These black bars are 32 mg / kg
At doses up to, papaverine alone did not induce catalepsy. More specifically,
Papaverine was administered at the indicated doses, either alone or with haloperidol (0.32 mg / kg), 30 minutes prior to testing. Each bar is the average latency to remove both forepaws from the lifted bar of six similarly treated animals. Kruskall-Wallace
Analysis of variance was used to compare the classified latencies of papaverine alone versus the addition of haloperidol. Post-hoc analysis indicates that animals receiving 3.2, 10, and 32 mg / kg of papaverine and haloperidol had significantly ( ^** ) longer latencies than animals treated with haloperidol alone .

FIG. 2 is two bar graphs showing the mean + SEM of the number of crossings of the animals during the first 60 minutes following administration of the substance, each in the shuttle box test. The top graph compares the effect of papaverine alone on exercise with the effect of papaverine on amphetamine-induced exercise. The graph at the bottom
B) shows the effect of papaverine alone on exercise by PCP
The effect of papaverine on induced movement is compared. Amphetamine is 1 mg / kg, i. p. Was administered. PCP was 3.2 mg / kg i.p. p. Was administered. Papaverine is administered at 32 mg / kg, i.p. p. Was co-administered with either drug. The data is n
-= Mean + SEM crossover number for the first 60 minutes following drug administration for 8 rats / group. ^** p <0.01 relative to vehicle / vehicle control; ^* p <0.05 relative to PCP by vehicle / t-test.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (Reference) A61P 25/22 A61P 25/22 25/24 25/24 25/30 25/30 25/32 25/32 25/36 25 / 36 43/00 111 43/00 111 (72) Inventor Lorraine Ann Level 06340, Connecticut, United States, Groton, Eastern Point Road, Pfizer Global Research and Development (72) Inventor Frank Samuel Mennity, Connecticut, United States 06340, Groton, Eastern Point Road, Pfizer Global Research and Development (72) Inventor Christopher Joseph Schmidt, Connecticut, United States 06340, Groton, Eastern Point Lord, Pfizer Global Research and Development

Claims (10)

[Claims] 1. A pharmaceutical composition for treating anxiety or psychiatric disorders in a mammal, comprising an effective amount of a selective PDE10 inhibitor for treating anxiety or psychiatric disorders in the mammal. 2. The mental disorder is, for example, paranoid, split,
Tension, undifferentiated, or residual schizophrenia; schizophrenic disorders; for example, delusional or depressed schizophrenic disorders; delusional disorders; substance-induced psychiatric disorders, such as alcohol, amphetamine, timer, cocaine; Psychosis induced by hallucinogens, inhalants, opioids, or phencyclidine; paranoid personality disorder; and schizophrenic personality disorder; and anxiety disorder is panic disorder; agoraphobia; 2. The pharmaceutical composition according to claim 1, wherein the composition is selected from: specific phobia; social phobia; obsessive-compulsive disorder; post-traumatic stress disorder; acute stress disorder; and general anxiety disorder. 3. A method for treating a dyskinesia in a mammal, comprising an effective amount of a selective PDE10 inhibitor for the treatment of a dyskinesia selected from Huntington's disease and dyskinesia associated with dopamine agonist treatment in the mammal. For pharmaceutical compositions. 4. A pharmaceutical composition for treating a movement disorder of a mammal, comprising an effective amount of a selective PDE10 inhibitor for treating a movement disorder selected from Parkinson's disease and lower limb anxiety syndrome in the mammal. 5. A drug addiction to a mammal, for example, alcohol,
A pharmaceutical composition for the treatment of said mammal comprising an effective amount of a selective PDE10 inhibitor for the treatment of amphetamine, cocaine, or opiate addiction. 6. A pharmaceutical composition for the treatment of a disorder in a mammal, comprising an effective amount of a selective PDE10 inhibitor in the treatment of a disorder in which cognition is impaired in the mammal. 7. The disorder according to claim 1, wherein the disorder is dementia, such as Alzheimer's disease, multiple infarct dementia, alcoholic dementia or other drug-related dementia, dementia associated with intracranial tumor or brain injury, Huntington's disease or Parkinson's disease. Nau dementia or AID
S-related dementia; delirium, amnestic disorder; post-traumatic stress disorder; mental retardation; learning disorders, such as reading, computing, or writing disorders; attention deficit / hyperfunction; 7. The method according to claim 6, wherein the cognitive decline is selected.
A pharmaceutical composition according to claim 1. 8. A pharmaceutical composition for treating a disorder or condition in a mammal, comprising an effective amount of a selective PDE10 inhibitor for treating the onset of a mood disorder or mood symptom in the mammal. 9. The mood disorder or mood symptom may be mild, moderate or severe, severe depressive symptom, manic or mixed mood symptom, or hypomanic mood symptom. Onset; onset of depressive mood symptoms of atypical signs; onset of depressive symptoms of depressive signs; onset of depressive symptoms of catatonic signs; onset of mood symptoms starting after parturition; Severe depressive disorder; dysthymia disorder; mild depressive disorder; premenstrual discomfort disorder; post-psychotic depressive disorder of schizophrenia; psychotic disorder such as delusional disorder or schizophrenia; 9. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition is selected from overlapping severe depressive disorders; manic depressive disorders, such as manic depressive I disorder, manic depressive II disorder, and circulatory disorders. 10. The pharmaceutical composition for selectively inhibiting PDE10 in a mammal, which comprises an effective amount of papaverine for selectively inhibiting PDE10 in the mammal.